REPORTS ters (14) containing normal Btbd3 expression (Fig. 2D). Total BTBD3 protein levels in somatosensory cortex did not differ between control and
NMDAR1 mutant mice (Fig. 2E). However, the subcellular localization of BTBD3 in wild-type brain partitioned to the nuclear fraction, whereas
Fig. 4. BTBD3 controls dendrite orientation in ferret visual cortex. (A) In situ hybridization of BTBD3 in the ferret brain at P16. (B) Dorsal view of P16 ferret brain that shows transfected neurons in the visual cortex (yellow arrows, left). The WGA-positive axon terminus is observed in the primary visual cortex (yellow bracket, middle). Positions of analyzed YFP-positive layer IV neurons are indicated by red arrows (right). (C) Reduction of BTBD3 mRNA by shRNA electroporation was tested by in situ hybridization (top, asterisks), and by quantitative real-time fluorescence polymerase chain reaction, (bottom). Lane 1, BTBD3 in control brain; lane 2, BTBD3 in shRNA electroporated knock-down brain; lane 3, B-ACTIN in control brain; lane www.sciencemag.org
SCIENCE
BTBD3 in mutant brain was found selectively in the cytosolic fraction (Fig. 2F). To study subcellular localization dynamics, we tagged Btbd3 with
4, B-ACTIN in shRNA electroporated knock-down brain. (D to F) Confocal images and schematic traces of neurons in control (D), 3 days after monocular enucleation (E) and monocular enucleation brain after BTBD3 knock-down (F). (G) Dendrite distribution was quantified by Sholl analysis, distribution of primary dendrites, and dendritic length. *P < 0.05, **P < 0.01, t test. (H) Dendrite morphology in control brain and monocular enucleation brain were observed at P51. (I) Dendrite distribution was quantified by Sholl analysis, distribution of primary dendrites, and dendritic length. *P < 0.05, **P < 0.01, t test. Scale bars are 1 mm in (A), 250 mm in (B) and (C), 25 mm in (D) to (F), and 50 mm in (H). VOL 342
29 NOVEMBER 2013
1117